Gilthead seabream (Sparus aurata) vitellogenin:: Purification, partial characterization, and validation of an enzyme-linked immunosorbent assay (ELISA)
Gilthead seabream (Sparus aurata) vitellogenin:: Purification, partial characterization, and validation of an enzyme-linked immunosorbent assay (ELISA)
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DOI：
10.1006/gcen.1998.7075
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发表时间：
1998-06-01
影响因子：
2.7
通讯作者：
Polzonetti-Magni, AM
中科院分区：
文献类型：
--
作者：
Mosconi, G;Carnevali, O;Polzonetti-Magni, AM
通过 DEAE-Sepharose 柱上的离子交换色谱法从鲷鱼血浆中纯化 VTG。对卵黄原蛋白进行了表征并测定了其性质。通过 Sephadex G-200 柱获得的天然形式的分子量约为 450 kDa，而在变性和还原条件下通过电泳检测到的表观分子量为 180 kDa，表明天然蛋白质为二聚体形式。使用刀豆球蛋白 A 测定碳水化合物的存在，而使用阳离子染色剂 Stains-all 检测磷酸基团的存在。这些数据与性别特异性、雌激素诱导性以及 abVTG 与主要卵黄蛋白的交叉反应性一起确定该蛋白为卵黄蛋白原。经过验证的 ELISA 可用于快速、可靠地测量雌性亲鱼中与雌二醇 17 β 相关的血浆 VTG 变化。 (C) 1998 年学术出版社。
VTG was purified from seabream Sparus aurata plasma by ion exchange chromatography on a DEAE-Sepharose column. The vitellogenin was characterized and its properties were determined. The molecular mass of the native form, obtained by Sephadex G-200 column, was around 450 kDa, whereas an apparent molecular mass of 180 kDa was detected by electrophoresis under denaturing and reducing conditions, suggesting a dimeric form for the native protein. The presence of carbohydrates was determined using concanavalin A, while the presence of phosphate groups was detected by Stains-all, a cationic stain. These data together with the sex specificity, the estrogen inducibility, and the cross-reactivity of the abVTG against the major yolk proteins identifies this protein as vitellogenin. The validated ELISA was used for a rapid and reliable measurement of plasma VTG changes related with those of estradiol-17 beta in female broodstock. (C) 1998 Academic Press.